Flowable matrix compositions and methods

ABSTRACT

Flowable matrix compositions and methods of their use and manufacture are provided. Exemplary compositions may include a flowable, syringeable, putty-like form of acellular human dermal matrix. In some cases, compositions may include a moldable acellular collagen extracellular matrix. In use, the matrix compositions can be used to fill or treat skin voids, channel wounds, and other soft tissue deficiencies.

CROSS REFERENCE TO RELATED APPLICATION

This application is a continuation application of U.S. patentapplication Ser. No. 13/712,295, filed Dec. 12, 2012, which claims thebenefit of the filing date of U.S. Prov. Patent Appl. No. 61/577,299,entitled “FLOWABLE MATRIX COMPOSITIONS AND METHODS,” filed Dec. 19,2011, the entire disclosures of which are incorporated herein byreference for all purposes.

BACKGROUND

Embodiments of the present invention are directed in general to thefield of wound treatments, and in particular to flowable human tissuecompositions, and methods of their use and manufacture.

Human tissue compositions, which may be derived from cadaveric donors,have been used for many years in various surgical procedures, includingtreatments for abrasions, lacerations, burns, and other wounds. Althoughhuman tissue compositions and methods are presently available andprovide real benefits to patients in need thereof, many advances maystill be made to provide improved dressing systems and methods fortreating patients. The flowable matrix compositions and treatment andmanufacture methods described herein provide further solutions andanswers to these outstanding needs.

BRIEF SUMMARY

Embodiments of the present invention provide flowable matrixcompositions and methods suitable for use in advanced wound management,including acute and chronic wound treatment, general surgery, plasticand reconstructive surgery, uro-gynecological surgery, and numeroussurgical procedures where an irregular tissue defect repair may bebeneficial.

An exemplary flowable matrix composition can be produced from a softtissue, such as an allograft human dermis which has been recovered froma tissue donor. The tissue can be aseptically processed to remove cells,while preserving natural biologic components and structural features ofthe tissue. The soft tissue can be further processed to yield aputty-like, pliable, matrix. The flowable matrix can be supplied forsurgical use as a sterile product. In some cases, the flowable matrixcan be loaded in an applicator, such as a syringe. An applicator orsyringe can be of any size or volume. For example, a 6 c.c. syringe maybe used. In use, a surgeon or other medical personnel may deliver theflowable matrix to an irregularly shaped wound site of a patient. Insome cases, a decellularized or partially decellularized matrix may beused for the repair or replacement of damaged or inadequateintegumentary tissue. In some instances, the soft tissue matrix isprepared from a tissue containing an amount of water or hydration. Insome cases, water may be added to the matrix during processing. In somecases, the preparation may be processed without the incorporation ofadditional water. In some cases, the matrix can be processed withoutremoving water therefrom. In some instances, the soft tissue matrixcomposition is at least partially hydrated when packaged. The surgeon ormedical professional may use the matrix composition directly uponopening the packaged product, without performing additional mixing orrehydration steps.

Flowable matrix embodiments of the present invention are well suited fora variety of therapeutic applications, including wound care and burncase. Exemplary wound care applications may involve treatment of chronicwounds, tunneling, channel, or invaginated wounds, wounds presenting adeep and irregular wound bed, and the like. The flowable, putty-like,compliant properties of the matrix render it particularly suitable forsuch indications. The flowable matrix or scaffold can be placed into thetunnels and tracts of chronic non-healing wounds which include, but arenot limited to, venous leg ulcers and diabetic foot ulcers, acute woundswhich include drained abscesses, and cysts. In some cases, the flowablematrix can be administered to a patient via a delivery cannula. In somecases, the flowable matrix can be delivered or syringed percutaneouslythrough a large gauge needle to the patient. In some cases, the flowablematrix can be administered to a patient as part of a surgical procedurefor treating a rotator cuff injury or tear.

In one aspect, embodiments of the present invention encompass methodsfor producing a soft tissue matrix composition for use in a patienttreatment. Exemplary production methods may include obtaining a portionof soft tissue material, and processing the portion of soft tissuematerial according to a protocol which comprises cryofracturing theportion of soft tissue material. The processing protocol may provide thesoft tissue matrix composition. In some instances, the soft tissuematerial is acellular. In some instances, the soft tissue material is atleast partially decellularized. In some instances, the soft tissuematerial is not partially or completely decellularized. According tosome embodiments, a processing protocol may include triturating thecryofractured soft tissue material. For example, a triturating step mayinclude milling a cryofractured portion of soft tissue material. In somecases, a processing protocol may include adding a wetting agent to thetriturated soft tissue material. In some cases, the wetting agent mayinclude a saline solution. In some cases, the soft tissue matrixcomposition may have a putty consistency. In some cases, the portion ofsoft tissue material is in a naturally hydrated state prior toprocessing according to the protocol. In some cases, the portion of softtissue material is in a partially hydrated state prior to processingaccording to the protocol. In some cases, the soft tissue material isnon-immunogenic or has reduced immunogenicity. In some cases, the softtissue material is reduced in cytotoxicity. In some cases, the softtissue material includes epidermal tissue, a dermal tissue, a placentalderived tissue, an amnion tissue, a chorionic tissue, a tendon tissue,an umbilical cord tissue, an intestine tissue, or a musculoskeletalnon-osseous tissue. In some cases, the soft tissue material includeshuman soft tissue, an equine soft tissue, a bovine soft tissue, aporcine soft tissue, an ovine soft tissue, a caprine soft tissue, or anavian soft tissue. In some cases, the soft tissue material includes ahuman dermal tissue. In some cases, the cryofracturing step includestreating the portion of soft tissue material with liquid nitrogen. Insome cases, the cryofracturing step includes treating the soft tissuematerial with liquid nitrogen for a period of about less than one hour.In some cases, the cryofracturing step includes treating the soft tissuematerial with liquid nitrogen for a period within a range from about 10second to about 1 minute. In some cases, the cryofracturing step rendersthe portion of soft tissue material stiff and friable. According to someembodiments, methods may include combining the soft tissue materialmatrix composition with a biocompatible carrier, a thickener, anadhesive, or any mixture, combination, or sub-combination thereof. Somemethods may include loading the soft tissue material matrix compositioninto an applicator assembly. According to some embodiments, anapplicator assembly may include a syringe mechanism or a cannulamechanism. In some cases, the cryofracturing partially disruptsextracellular matrix tissue macro architecture within the soft tissuematerial. In some cases, the cryofracturing step partially disruptsextracellular matrix tissue macro architecture within the soft tissuematerial, and the triturating step further disrupts extracellular matrixtissue macro architecture within the soft tissue material. In somecases, the soft tissue material includes a delaminated dermal tissue. Insome cases, the soft tissue material includes a non-delaminated dermaltissue. In some cases, methods may include combining the soft tissuematrix composition with a stromal volume fraction, a progenitor cellpopulation, or a stem cell population.

In another aspect, embodiments of the present invention encompassmethods for producing a soft tissue matrix composition for use in apatient treatment. Exemplary methods may include obtaining a portion ofsoft tissue material, and partially homogenizing the portion of softtissue material so as to produce the soft tissue matrix composition.

In another aspect, embodiments of the present invention encompass softtissue matrix compositions for use in a patient treatment. Exemplarycompositions may include a soft tissue material having a mechanicallydisrupted macrostructure and a partially disrupted collagenmicrofibrillar architecture. In some cases, a soft tissue matrixcomposition may include a wetting agent. In some cases, the wettingagent may include a saline solution. In some cases, the composition ispresent in an applicator assembly. In some cases, the soft tissuematerial is acellular. In some cases, the soft tissue material is atleast partially decellularized. In some cases, the soft tissue materialis not partially or completely decellularized. In some cases, the softtissue matrix composition does not adhere to a surgical glove material.In some cases, the surgical glove material comprises a member selectedfrom the group consisting of latex, neoprene, vinyl, and Nitrile. Insome cases, a matrix composition may also include a stromal volumefraction, a progenitor cell population, or a stem cell population.

In still a further aspect, embodiments of the present inventionencompass methods for treating a soft tissue of a patient. Exemplarymethods may include obtaining a soft tissue matrix composition, andadministering the soft tissue matrix to the soft tissue of the patient.In some cases, the soft tissue material is acellular. In some cases, thesoft tissue comprises a defect. In some cases, the soft tissue defect isa skin void, and an administering step includes at least partiallyfilling the skin void with the soft tissue matrix composition. In somecases, an administering step includes delivering the soft tissue matrixwith an applicator assembly to the soft tissue, and the applicatorassembly includes a cannula mechanism or a syringe mechanism. In somecases, an administering step includes manually delivering the softtissue matrix with a gloved hand. In some cases, the soft tissue defectis a skin void, a channel wound, an ulcer, a surgical wound, a traumawound, a chronic wound, an acute wound, or an exsanguinating site. Insome cases, the soft tissue defect is an ulcer wound such as a pressureulcer, a venous ulcer, a diabetic ulcer, or a chronic vascular ulcer. Insome cases, the soft tissue defect is a surgical wound such as a generalsurgery wound, a plastic surgery wound, a reconstructive surgery wound,a urological surgery wound, or a gynecological surgery wound. In somecases, methods may also include at least partially covering the matrix,the tissue, or both, with a wound dressing. In some cases, methods mayalso include forming the soft tissue matrix composition into a shapesuitable for placement at or within the soft tissue. In some cases, thesoft tissue defect is an exsanguinating site, and administration of thesoft tissue matrix activates a hemostatic cascade at the exsanguinatingsite. In some cases, the soft tissue matrix composition also includes astromal volume fraction, a progenitor cell population, or a stem cellpopulation.

The above described and many other features and attendant advantages ofembodiments of the present invention will become apparent and furtherunderstood by reference to the following detailed description whenconsidered in conjunction with the accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates aspects of a method for producing a soft tissuematrix composition according to embodiments of the present invention.

FIG. 2 illustrates aspects of a method for producing a soft tissuematrix composition according to embodiments of the present invention.

FIG. 3 illustrates aspects of a method for producing a soft tissuematrix composition according to embodiments of the present invention.

FIG. 4 illustrates aspects of a method for treating a soft tissue of apatient according to embodiments of the present invention.

FIG. 5 shows aspects of a process for administering a soft tissue matrixcomposition to a patient, according to embodiments of the presentinvention.

FIG. 6 shows aspects of a process for administering a soft tissue matrixcomposition to a patient, according to embodiments of the presentinvention.

FIG. 7 shows aspects of an exemplary flowable matrix according toembodiments of the present invention.

FIG. 8 shows aspects of an exemplary flowable matrix according toembodiments of the present invention.

FIG. 9 shows aspects of an exemplary flowable matrix according toembodiments of the present invention.

DETAILED DESCRIPTION

Embodiments of the present invention encompass flowable matrixcompositions and methods. Exemplary compositions may include a flowable,putty-like form of acellular human dermal matrix that can be deliveredfrom a syringe. In some cases, compositions may include a moldableacellular collagen extracellular matrix. In use, the matrix compositionscan be used to fill or treat skin voids, channel wounds, and other softtissue deficiencies.

Material composition embodiments of the invention may include anacellular human dermal matrix (e.g. a collagenous matrix derived fromfull-thickness or partial-thickness human skin that has been subjectedto a gentle decellularization process) and a moistening solution such asnormal saline, phosphate buffered saline, Lacted Ringers solution, or asimilar physiological compatible moistening solution. In some cases, thematerial may include only the dermal matrix and the moistening solution.

According to some embodiments, a composition may include any suitablebiocompatible extracellular matrix material derived from humans,animals, sources such as allograft material (human derived tissue),placental material, xenograft material including bovine, ovine, caprine,porcine, equine, avian, or other animal sourced tissue. In addition tothese materials, matrix compositions may include any collagenousmaterial capable of being delivered through a cannula. In addition tohuman tissue allograft compositions for transplantation, other materialsas noted herein can be used, for transplantation and other therapeuticpurposes.

Flowable matrix compositions, such as a human collagenous matrix, can beused in the surgical fields, for example where it may be desirable tofill or treat an acute incisional defect or chronic wound with thematrix for the purpose of allowing cellular infiltration andangiogenesis to occur in order to affect a post-surgical healingresponse. In some cases, a primary medical indications for use mayinclude the management of wounds or conditions, including withoutlimitation partial and full-thickness wounds, pressure ulcers, venousulcers, diabetic ulcers, chronic vascular ulcers, tunneled/underminedwounds, surgical wounds, donor sites/grafts, post-Mohs surgery, postlaser surgery, podiatric, wound dehiscence, and trauma wounds such asabrasions, lacerations, second-degree burns, and skin tears. In someembodiments, a matrix composition may be intended for a one-time use.

Relatedly, flowable matrix compositions may be use in the therapeuticintervention of chronic and acute wounds. In some cases, matrixcompositions may permit or facilitate the in situ delivery of ahemostatic material to an exsanguinating site. In some cases, a flowablematrix composition may provide a vehicle for the delivery of collagen,as a hemostatic material, to hard to reach bleeding sites. The collagencan operate to activate the hemostatic cascade by the mechanism ofplatelet aggregation. Subsequent to platelet adhesion and aggregationinitiated by collagen, the hemostatic effect of fibrinogen beingactivated to form a fibrin clot ensues thereby forming a clot anddiminishing or stopping the flow of blood from the injured site.

In clinical applications, matrix compositions can be used for softtissue integumental repair. For example, a putty-like, moldableacellular dermal matrix can be delivered through a cannula or othersyringe attachment to a treatment site. In some cases, matrixcompositions can be used to deliver a hemostatic material (e.g.collagen) to a bleeding site. Matrix compositions can be used to treatchanneling wounds, deep full-thickness integumental defects, and otherwounds or conditions where a putty-like, moldable matrix is suitable forapplication. In some case, matrix compositions provide a beneficialtherapeutic use for achieving hemostasis at a bleeding site.

Embodiments of the present invention encompass matrix compositions whichcan be molded or otherwise handled by a gloved hand without sticking tothe glove. Relatedly, exemplary matrix compositions can be used withoutsticking to operative instruments which may come into contact with thematrix. Hence, matrix compositions can be easily applied with a glovedhand without sticking to the gloves or other surfaces.

According to some embodiments, matrix compositions can be applied to asoft tissue defect such as a channeling wound in similar fashion to a“putty-like” material wherein a medical practitioner may mold a portionof the matrix composition into an irregularly shaped soft tissue voidwith a gloved finger. In some cases, a syringe equipped with a standardmedical cannula of appropriate length can be used to deliver the matrixcomposition to a treatment site. Relatedly, in some cases a matrixcomposition can be supplied in pre-loaded syringe assembly. Optionally,a surgeon or practitioner may use the syringe and cannula as a directeddelivery device whereby the matrix composition is delivered to anirregular or channel wound site or other soft tissue defect.

Matrix compositions can be applied to a patient or individual so as tofill irregular wounds or soft tissue defects in a complete fashion,which may otherwise be difficult to treat with rigid flat or rectangulardevices. By providing a flowable material which completely contacts awound bed, improvement in healing can be realized. Such enhanced orcomplete contact with the wound bed promotes proper revascularization,angiogenesis, and wound healing. Matrix compositions are particularlywell suited for use in treating an irregularly shaped wound bed, due tothe enhanced contact between the matrix and the wound.

According to some embodiments, a full-thickness human skin can betreated to remove the epidermal layer, and optionally further treated toat least partially remove the cellular contents and nucleic detritus.The resultant human dermal matrix, optionally in acellular form or atleast partially in acellular form, can then be treated by acryofracturing technique to render it stiff and friable. The friablematrix can then be triturated, for example by milling. The resultantcomposition matrix material can provide a putty-like consistency,optionally upon thawing. In some cases, the matrix composition is not ina powder form, and hence is not as susceptible to the build up of staticelectrical charges. According to some embodiments, the soft tissuematrix composition is provided to the surgeon or administered to thepatient in a native form, without the presence of other materials oradditives.

In some cases, a matrix composition, such as a flowable collagenousmatrix material, may include various biocompatible carriers, thickeners,or adhesives, including but not limited to carboxymethylcellulose,poloxymer, and fibrin sealant. Likewise, the friable matrix can betriturated by any of a variety of triturative methodologies, such aspartial homogenization of a non-cryotreated acellular matrix andvariations thereof. Such triturations can be performed so as to provideparticular matrix particle sizes, in some instances.

Turning now to the drawings, FIG. 1 illustrates a method of producing asoft tissue matrix composition for use in a patient treatment, accordingto embodiments of the present invention. Manufacturing method 100includes obtaining a portion of soft tissue material, as shown by step110, processing the portion of soft tissue material according to aprotocol which includes cryofracturing the portion of soft tissuematerial, as shown by step 120. The processing protocol provides thesoft tissue matrix composition, as shown by step 130. In some cases, thesoft tissue material is acellular, or at least partially acellular. Forexample, the soft tissue material may be processed according to adecellularization treatment to remove some or all of the cellularcomponents therefrom. Decellularization may be accomplished by knowntechniques such as those referenced in Gilbert, TW, Sellaro, TL andBadylak, SF; “Decellularization of tissues and organs”, Biomaterials,27, (2006), 3675-3683, the content of which is incorporated herein byreference. Embodiments of the present invention also encompass tissuewhich are processed according to decellularization techniques such asthose described in U.S. Pat. No. 5,336,616, the content of which isincorporated herein by reference. In some cases, the processing protocolof step 120 includes triturating the cryofractured soft tissue material.Optionally, the protocol of step 120 may include adding a wetting agentto the triturated soft tissue material. In some cases, the wetting agentincludes a saline solution. In some cases, a wetting agent may includeany of the known surface active amphiphilic chemicals that typically actto lower the surface tension of aqueous solvents and are capable offorming micelles. Such wetting agents may be any of the surfactantsknown in the field as anionic, cationic or nonionic surfactants. Anonexhaustive list of specific wetting agents includes phosphatebuffered saline (PBS), sodium dodecyl sulfate, sodium stearate,benzalkonium chloride,3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS),lecithin, TRITON® X-100, and Nonoxynol-9. According to exemplaryembodiments of the present invention, the soft tissue matrix compositionhas a putty consistency or viscosity. Relatedly, the soft tissue matrixcomposition may have a shear resistance force within a range from about250 Pascal seconds (Pa s) to about 2500 Pascal seconds. In some cases,the soft tissue matrix composition has a shear resistance force within arange from about 1000 Pascal seconds to about 2000 Pascal seconds. Suchshear resistance forces or consistencies can be measured by standardviscometric or rheometric methods, including without limitationBrookfield viscometers, Stormer viscometers, ICI Cone and Plateviscometers, rheometers, and the like. In some instances, the shearresistance or flowability of the matrix composition can be modified byadjusting an amount of wetting agent added to the triturated soft tissuematerial. In some cases, a matrix composition can be produced withoutincluding a wetting agent. Such matrix compositions may present a moreputty-like consistency (e.g. higher viscosity), as compared with matrixcompositions which include a wetting agent (e.g., lower viscosity). Theamount of wetting agent may determine which types of applicationprocesses can be used to administer the composition. For example, ahighly flowable matrix composition containing a larger amount of wettingagent may be suitable for delivery by a syringe, and a less flowablematrix composition containing little or no wetting agent may be suitablefor manual delivery as a molded or formed plug or filling. In somecases, a plug can be manually formed. In some cases, a plug can bemechanically formed, for example with a press mechanism. Optionally, aplug can be molded into a desired shape using both manual and mechanicalforming techniques. In some cases, a plug can be formed so that itprovides a shape or surface that is complimentary to a shape or surfaceof a patient wound. For example, for treating a patient wound presentinga cylindrical channel feature, the plug can be formed into a shapepresenting a complimentary tubular feature which is configured forplacement in the patient wound channel feature.

In some cases, an exemplary flowable matrix preparation may bedeliverable or flowable through an 18 g needle (1.27 mm; 0.050 inches)but not a 26 g needle (0.4636 mm; 0.01825 inches).

According to some embodiments, the portion of soft tissue material maybe in a naturally hydrated state prior to processing according to theprotocol of step 120. In some instances, the portion of soft tissuematerial may be in a partially hydrated state prior to processingaccording to the protocol of step 120. In some instances, the processingprotocol of step 120 may include triturating a cryofractured soft tissuematerial. Trituration may be accomplished by blending or millingtechniques, for example. In some instances, a blender, mill, or othertrituration process may be selected or performed so as to provide matrixparticles of various sizes. Relatedly, different administrationtechniques may involve the delivery of different particle sizes. Forexample, the particle size of the of the matrix can be increased ordecreased to allow the product to be delivered through a cannula (e.g.larger particles) or through a needle (e.g. smaller particles),respectively.

According to some embodiments, the matrix composition presents aconsistency or degree of flowability, such that it can be easilydispensed or extruded through syringe opening (e.g 3 mm orifice) or acatheter or tube (e.g. inner diameter less than 4 mm) upon applicationof a standard amount of syringe pressure which may be applied manuallyin the surgical setting.

In some cases, the soft tissue material or matrix composition isnon-immunogenic, or has reduced immunogenicity in comparison to a tissuematerial which has not been processed according to embodiments describedherein. For example, the material or composition may elicit no immuneresponse, or a limited immune response, when introduced into a patient.In some cases, the soft tissue material or matrix composition isnon-cytotoxic, or has reduced cytotoxicity in comparison with a tissuematerial which has not been processed according to embodiments describedherein. Techniques for the decellularization processes may entail theuse of detergents, surfactants, acids, bases, salts, and other chemicalentities which result in the presence of detectable residual chemicalsin the acellular tissue matrix. According to some embodiments, it may bedesirable to sufficiently remove these residuals, which may otherwise insome cases exhibit a cytotoxic response if they still remain in certainamounts following decellularization. In this way, removal of theresiduals can reduce the possibility of a cytotoxic response. In somecases, such cytotoxicity may prevent, stop, or otherwise inhibitsuccessful incorporation of the matrix into the implanted host.Therefore it may be desirable to remove as much of any residualdecellularizing process chemicals as possible. According to someembodiments, the trituration or comminution of the tissue followed bysuccessive washing steps can maximize the surface area of the tissueparticles that are exposed to the washing solution, therebysignificantly reducing the presence of any potential residual processchemicals. This significant reduction of residuals will minimize orprevent a potentially cytotoxic response of the final matrix product.Such washing agents may include any of the several solvents that aremiscible with a hydrated medium and may exhibit hydrophobic orhydrophilic solvation characteristics depending upon thedecellularization agent used and may be exemplified by sterile water,sterile physiological saline, sterile phosphate buffered saline, or anynumber of mixtures of these with organic solvents such as 1-propanol,2-propanol, acetone, methanol or similar minimally polar solvents.

Soft tissue material, such as the material obtained in step 110, mayinclude an epidermal tissue, a dermal tissue (e.g. full-thickness orpartial-thickness), a placental derived tissue, an amnion tissue, achorionic tissue, a tendon tissue, an umbilical cord tissue, anintestine tissue, a musculoskeletal non-osseous tissue, or the like. Insome cases, soft tissue material may include a human soft tissue, anequine soft tissue, a bovine soft tissue, a porcine soft tissue, anovine soft tissue, a caprine soft tissue, an avian soft tissue, or thelike. According to exemplary embodiments, the soft tissue materialincludes a human dermal tissue. For example, the soft tissue materialmay include full-thickness or partial-thickness human skin. In somecases, soft tissue material may include a cartilage tissue or a muscletissue.

The processing protocol illustrated in step 120 may include subjectingthe tissue material to a cryofracturing technique. For example, step 120may include treating the portion of soft tissue material with liquidnitrogen. In some cases, a cryofracturing step may include treating thesoft tissue material with liquid nitrogen for a period of about lessthan one hour. In some cases, the cryofracturing step may includetreating the soft tissue material with liquid nitrogen for a periodwithin a range from about 10 second to about 1 minute. According toexemplary embodiments, the cryofracturing step may render the portion ofsoft tissue material stiff and friable. For example, cryofracturingprocesses, which may include immersion in liquid nitrogen that appliedto a soft tissue material can render it suitably friable for subsequenttrituration. A soft tissue material matrix composition, such as thatobtained in step 130, may be combined with a biocompatible carrier, athickener, or an adhesive. Exemplary carriers or thickeners may includeany of the Generally Recognized as Safe (GRAS) excipients such ascarboxy methyl cellulose (CMC), methylcellulose, agar, carrageenan,modified starch, pectin, poloximers or similar materials. Optionally,adhesives such as fibrin glue may be employed. Such fibrin glue can bederived from recombinant human fibrin combined with human or bovinederived thrombin. A soft tissue matrix composition may also be loadedinto an applicator assembly, for subsequent use by a surgeon or othermedical personnel. In some cases, an applicator assembly may include asyringe mechanism and a cannula mechanism.

The processing protocol of step 120 may include cryofracturing the softtissue material so as to at least partially physically disruptextracellular matrix collagen or tissue macro architecture within thesoft tissue material. In some cases, as further discussed elsewhereherein, a cryofracturing process may partially disrupt extracellularmatrix collagen or tissue macro architecture within the soft tissuematerial, and a subsequent triturating process may further disruptextracellular matrix collagen bundles within the soft tissue material.

Either or both of the cryofracturing and triturating processes may beapplied to any of a variety of tissue types, including withoutlimitation amnion tissue, umbilical cord tissue, intestine tissue, orother soft tissues. Any of these tissues may be similarly treated eitherwith or without a prior decellularization treatment. Hence, for example,skin, amnion, tendon, or other tissues can be cryofractured and renderedinto a putty-like medium. In some embodiments, such tissues may berendered non-immunogenic prior to manufacturing them into a putty, forexample if the end use is implantation within a patient. According tosome embodiments, a decellularization treatment may render the tissuenon-immunogenic. In some cases, a decellularization treatment may atleast partially reduce or eliminate immunogenicity in the tissue. Hence,where a matrix material is intended for use in a topical treatment,embodiments may involve at least partially decellularizing the tissue,or processing the tissue without performing a decellularization step.Relatedly, where a matrix material is intended for use in animplantation procedure, embodiments may involve at least partially orfully decellularizing the tissue. In other words, i the intended medicaluse for the soft tissue matrix composition is that of a temporarycovering or product, and not a permanent implantation, the matrix may besuitable for use after being subjected to a partial decellularizationregimen or no decellularization regime at all. In some cases, thedecellularization techniques can be applied to any of a variety of humanand/or heterograft derived matrices.

According to some embodiments, a stromal vascular fraction (SVF) fromadipose tissue (see e.g. Gimble et al.; Circulation Research. 2007; 100:1249-1260 , the contents of which are incorporated herein by reference),or other compositions containing progenitor cell populations, stem cellpopulations, or mixed populations thereof, may be added to the softtissue matrix. Such fractions or populations may be obtained fromcadaveric human tissue or other suitable sources. In some instances,when added to the flowable matrix composition, mesenchymal stem cellsmay adhere to the flowable matrix substrate. Exemplary techniques forobtaining stromal vascular fractions, progenitor cells, and stem cellsfrom tissue are described in PCT Publication WO 2010/059565 to Shi, thecontents of which are incorporated herein by reference.

In some cases, the soft tissue material may be a delaminated dermaltissue. In some cases, the soft tissue material may be a non-delaminateddermal tissue. For example, dermal tissue may have an intact epidermallayer. Alternatively, dermal tissue may be treated to remove theepidermal layer, either partially or completely. Delamination ordeepidermization may be accomplished by any of the typical methods knownin the field including mechanical debridement of the epidermis byrepeated scraping with a sharp instrument such as a surgical scalpel.Alternatively deepidermization may be chemically induced by any of thehypotonic or hypertonic saline methods such as Wilsteed, E M et al.; “Anultrastructural comparison of dermo-epidermal separation techniques”, J.Cutan. Pathol. 18: 8-12 (1991), the content of which is incorporatedherein by reference.

According to some embodiments, a soft tissue matrix composition may beproduced without treating the source soft tissue material with acryofracturing protocol. FIG. 2 illustrates aspects of a method 200 forproducing a soft tissue matrix composition for use in a patienttreatment. As shown here, method 200 includes obtaining a portion ofsoft tissue material, as depicted by step 210, partially homogenizingthe portion of soft tissue material, as depicted by step 220, so as toproduce the soft tissue matrix composition, as depicted by step 230.

FIG. 3 illustrates aspects of matrix composition manufacturing methods,according to embodiments of the present invention. As shown here,manufacturing method 300 may include treating a portion of soft tissuematerial with a decellularization process, as indicated by step 310.Decellularization may be accomplished by known techniques such as thosereferenced in Gilbert, T W, Sellaro, T L and Badylak, S F;“Decellularization of tissues and organs”, Biomaterials, 27 , (2006),3675-3683 , the content of which is incorporated herein by reference.Embodiments of the present invention also encompass tissue which isprocessed according to decellularization techniques such as thosedescribed in U.S. Pat. No. 5,336,616 , the content of which isincorporated herein by reference. In some cases, the soft tissuematerial may be completely decellularized. In some cases, the softtissue materially may be partially decellularized. Optionally, the softtissue material may not be subjected to a decellularization process.Following the decellularization treatment, the soft tissue material maybe cut, minced, or sliced into pieces, as indicated by step 320. Forexample, a portion of decellularized soft tissue material may be cut orseparated into small strips or pieces. Optionally, a wetting agent suchas sterile saline can be admixed with the minced pieces, for exampleafter the pieces have been minced in a blender, at which point thetissue material may be the consistency of a putty. In some instances,the cutting process may be performed for about 5 minutes. In someinstances, the duration of this time period may vary depending on thetype of tissue processed. In some instances, following adecellularization treatment, tissue or material can be processed toremove potentially antigenic cellular debris and proteinaceous materialsfrom the ECM.

Decellularized tissue material can be subjected to a cryo-hardeningtreatment, as indicated by step 330. For example, the tissue may beimmersed in liquid nitrogen, so as to freeze or cryofractured thetissue. In some cases, the soft tissue is treated with liquid nitrogenfor a duration of about one minute, or longer. As depicted by step 340,the frozen or cryo-treated soft tissue material may be triturated in ablender. In some instances, the tissue is processed in the blender for aduration of about one minute. In some instances, a wetting agent, or anyother additive, may be added to the soft tissue material before, during,or after the trituration process. Exemplary additives may include any ofthe pharmaceutical excipients that are classified by FDA as generallyrecognized as safe (GRAS). In some instances, the wetting agent is a0.9% sterile saline solution. As depicted in step 350, the trituratedsoft tissue material may then be placed or loaded into a filler syringe.Thereafter, as depicted in step 360, the soft tissue material may beplaced or loaded into applicator syringes, for example by extruding thesoft tissue material from a filler syringe into an applicator syringe.The loaded applicator syringes can then be packaged, as indicated bystep 370. Subsequently, the packaged applicator syringes can besterilized, for example by exposure to e-beam. According to someembodiments, a 48 cm² amount of decellularized soft tissue may yieldapproximately 10 c.c. to 15 c.c. of flowable matrix composition, whichmay in some instances provide enough product to load two or threesyringes. The entire process shown in FIG. 3 may take approximately 30minutes or less, in some instances.

FIG. 4 illustrates aspects of a therapeutic treatment according toembodiments of the present invention. A method 400 for treating apatient may include obtaining a soft tissue matrix composition, asdepicted by step 410, and administering the soft tissue matrix to thesoft tissue of the patient, as depicted by step 420, so as to treat thesoft tissue of the patient, as depicted by step 430. A patient may be ahuman or non-human animal. In some cases, the treated soft tissue of thepatient referred to in step 430 includes a soft tissue defect.Relatedly, a soft tissue defect may be present in the patient as a skinvoid, and the administering protocol of step 420 may include at leastpartially filling the skin void with the soft tissue matrix composition.In some cases, administration step 420 may include delivering the softtissue matrix with an applicator assembly to the soft tissue. Anapplicator assembly may include a cannula mechanism, a syringemechanism, or the like. In some cases, administration step 420 mayinclude manually delivering the soft tissue matrix with a gloved hand.For example, a surgeon may manually administer the soft tissue matrix tothe patient tissue or treatment site. According to some embodiments ofthe present invention, a soft tissue defect may be or include a skinvoid, a channel wound, an ulcer, a surgical wound, a trauma wound, achronic wound, an acute wound, an exsanguinating site, or the like.According to some embodiments of the present invention, a soft tissuedefect may be or include an ulcer wound, such as a pressure ulcer, avenous ulcer, a diabetic ulcer, or a chronic vascular ulcer. Accordingto some embodiments of the present invention, a soft tissue defect maybe or include a surgical wound, such as a general surgery wound, aplastic surgery wound, a reconstructive surgery wound, a urologicalsurgery wound, or a gynecological surgery wound. Some treatment methodsmay include at least partially covering the matrix, the tissue, or both,with a wound dressing. Some treatment methods may include molding orforming the soft tissue matrix composition into a shape suitable forplacement at or within the soft tissue. Such administration techniquescan be performed by a surgeon or other suitable medical personnel. Insome cases, a soft tissue defect may include an exsanguinating site, andadministration of the soft tissue matrix can operate to activate ahemostatic cascade at the exsanguinating site.

According to some embodiments, a flowable acellular human dermal matrixmaterial can be prepared and administered as follows. First, human skincan be recovered in a cutting operation. For example, humanfull-thickness skin can be severed from a donor, and cut intoappropriately sized pieces. The skin can then be processes according toselected delamination, decellularization, or washing steps, orcombinations thereof. Subsequently, the skin can be processed accordingto a cryofracturing procedure, which may involve exposing the skin toliquid nitrogen. Thereafter, the cryofractured skin can be milled ortriturated. The resulting putty-like collagen matrix can then be loadedinto a syringe for subsequent packaging and sterilization. An exemplaryproduct may be provided as 5 cc syringe loaded with approximately 3 c.c.(or more) of the soft tissue matrix composition. The loaded syringe canbe packaged in a TYVEK® peel inner clear plastic tub. This inner tub canbe placed in a TYVEK® peel outer tub. The resulting package can besterilized by e-beam and provided in a double sealed configuration foroperating room (OR) use. The flowable matrix product may be indicatedfor replacement or repair of damaged or inadequate integumental tissue.

As shown in FIG. 5, the flowable matrix composition can be syringedthrough a cannula into a skin void or tissue wound of a patient. In someembodiments, a tissue treatment product may include a 6 c.c. syringecontaining 3 c.c. of flowable matrix composition. In some embodiments, atissue treatment product may include a 6 c.c. syringe containing 5 c.c.of flowable matrix composition. In use, the flowable matrix compositioncan be applied to any of a variety of wound types, such as tunnelingwounds, channel wounds, invaginated wounds, and the like. According tosome embodiments, the matrix composition has a consistency such that itcan be delivered or extruded through a standard syringe (e.g. 3 mmorifice) or a cannula, catheter, or similar tube having an innerdiameter of less than 4 mm. As shown here, the matrix composition can bedelivered from a syringe into a wound bed that has an irregular shape orthat is difficult to access. In some embodiments, the matrix compositionmay be applied manually, for example as a putty. The matrix compositiontypically does not adhere to surgical glove material. Hence, theflowable product can be easily molded and pressed or formed into patientwounds, without sticking to the surgeon's gloved hand or finger. Asshown in FIG. 6, after the matrix composition is syringed through acannula and into a tissue void, the surgeon or other medical personnelcan cover the matrix composition and the patient defect with a wounddressing.

With a flowable matrix composition, embodiments of the present inventionprovide therapeutic interventions that allow for complete contact of thematrix with an irregular or channeling wound bed. Such materials can bemolded or formed, and placed in an irregular wound bed so thatsufficient material coats the wound bed to allow for a revascularizationand remodeling response to occur. Hence, embodiments provide a suitablematrix for facilitating a healing response. In some instances, matrixcompositions of the present invention provide a flowable acellular humandermal matrix, suitable for tissue regeneration indications.

According to some embodiments, soft tissue material can include astructural entity referred to as the extracellular matrix (ECM). The ECMmay surround and support cells which are located within the tissue.Often, the ECM may include collagen protein, present in a triple helixfibrillar configuration. Collagen fibers can provide tensile strengthand elasticity to the structure of the ECM. During processing proceduresas described herein, the macrostructure of the soft tissue, such as thehuman dermis, can be mechanically disrupted. It has been discovered thatalthough the integrity of the tissue extracellular matrix may becompromised during certain processing procedures (e.g. cryofractured,trituration), the microstructure of the matrix composition can remainintact, or sufficiently intact, for treatment purposes. Relatedly,according to some embodiments, a cryofracturing step can partiallydisrupt extracellular matrix collagen bundles within the soft tissuematerial, and a triturating step can further disrupt extracellularmatrix collagen bundles within the soft tissue material. Yet, themicrofibrillar architecture or structure of the collagen remainssufficiently intact and is available for tissue regeneration. Accordingto some embodiments, the terms microstructure and microarchitecture maybe used interchangeably. Hence, the disrupted microstructure of thecollagen may also be referred to as a disrupted or disturbedmicroarchitecture. The microstructure may be measured by histologicalanalysis. For example, tissue material at various stages of theprocessing (e.g. as shown in FIG. 3), can be histologically stained andmicroscopically observed. In some instances, the tissue material may bestained with Masson's Trichrome (MT) stain. Such stains can help tovisualize the microarchitecture of the collagen bundles that mainly formthe extracellular matrix. According to some embodiments, some amount ofphysical disruption may occur during the deep freeze or cryofracturedstep, and additional or more pronounced disruption may occur during thetrituration/comminution (e.g., blender) steps. The fast freezing oftissue which occurs at liquid nitrogen temperatures may causeinterstitial water within the tissue to freeze rapidly. Without beingbound by any particular theory, it is thought that rapidly forming icecrystals may exert expanding pressure to somewhat disrupt themicroarchitecture. Subsequent exposure to the blender step can furthercomminute the partially disrupted microarchitecture or microfragments.

EXPERIMENTAL EXAMPLE 1

A single piece of decellularized human dermal tissue, prior toundergoing a sterilization process, was stored in liquid Nitrogen for aperiod of time to render the tissue friable. The frozen tissue wasmilled. The resulting matrix composition, which had the consistency of aputty or paste, was placed into a 5 cc syringe by means of a spatula.The plunger on the syringe was depressed upon the paste to minimize airpockets within the matrix composition. The loaded syringe was placed ina plastic inner tub and heat-sealed with a first TYVEK® cover. Thisassembly was placed in a larger outer tub, and heat-sealed with a secondTYVEK® cover. The entire resulting syringe and package assembly wassubmitted to approximately 22 kGy electron beam radiation for sterilityassurance. Upon subsequent evaluation of the soft tissue matrixcomposition, it was observed that the matrix composition providedexcellent handling and ease of use characteristics.

EXPERIMENTAL EXAMPLE 2

Six pieces of decellularized (donor derived full-thickness dermaltissue) were obtained (4 cm×8 cm in size). The pieces were each cut intolong strips with a knife, and then frozen in liquid nitrogen. The frozenstrips were processed in a 200 ml Waring blender. At the onset ofblending, loud cracking emanated from the blender. Blending wascontinued until the loud cracking sounds ceased. 10 g of blended tissuewas combined with 1.5 g of 0.9% saline, and the combination was mixedwith a spatula, so as to form a matrix composition. The mixed matrixcomposition was loaded into an irrigation syringe having needle with a 3mm bore. Upon depression of the syringe plunger, the matrix compositionwas observed to flow easily through the 3 mm bore needle. Uponadditional testing, it was observed that the matrix composition flowedeasily through needles having bore sizes of 3 mm and 1.5 mm. However,the matrix composition was not observed to flow easily through an 18gauge needle. Five samples, each containing approximately 5 ml of matrixcomposition, were prepared for delivery to an e-beam sterilizationfacility.

EXPERIMENTAL EXAMPLE 3

Flow characteristics of skin, tendon, and decellularized dermal matrixmaterials were evaluated. Samples of each type were prepared, and loadedonto a syringe. The produce was delivered to a luer-loc tipped syringeonto which a luer lock needle could be attached securely. Each samplewas loaded along with an amount of 0.9% saline. Each of the threesamples were extrudable through a 16 gauge needle, but not a 23 gaugeneedle. A 1 cc sample of each material was diluted and dissolved in 9 ccof 0.9% saline, respectively. Each of the resulting 10 cc compositionswere extrudable through an 18 gauge needle (1.27 mm; 0.050 inches), butnot a 23 gauge needle (0.4636 mm; 0.01825 inches). Each of the threesample types were subject to 25-35 kGy E-beam sterilization, and aftersterilization the extrudability of the contents in the syringe werecompared to a sample that had not been e-beam sterilized. All threesterilized samples showed no difference in the ease of extrusion fromthe syringe with a 3 mm orifice.

EXPERIMENTAL EXAMPLE 4

FIGS. 7 and 8 illustrate 10× and 20× magnifications, respectively, ofexemplary flowable matrix embodiments stained with Masson's Trichromestain. MT stain is often used by histopathologists to visualizeconnective tissue in a material. Connective tissue of interest in theinstant application may primarily be made up of collagen. MT stainscollagen various hues of blue. Hence, FIGS. 7 and 8 show a predominantlycollagenous matrix derived from a decellularized full thickness humanskin source. The collagen matrix in a normal (i.e. non-triturated)extracellular matrix is usually “rope-like” (i.e. a tightly boundfibrous structure). For example, FIG. 9 shows a typicalnon-cryofractured extracellular matrix structure prior to a triturationstep. This Masson's Trichrome stain shows the collagenousmicroarchitecture as intact as opposed to comminuted. An unexpected andsurprising result of a cryofracturing process is the comminutedappearance of the material in FIGS. 7 and 8. More specifically, onewould expect a fibrous structure of FIG. 9 to simply be broken, cut ,orotherwise reduced into much smaller “ropes” or fibers followingtrituration. However the material shown in FIGS. 7 and 8 appears to havebeen comminuted (i.e. pulverized or otherwise reduced to particles).Although a portion of the flowable matrix depicted in FIGS. 7 and 8 hasa few identifiable fibers, the majority of it is in a comminuted form.

Delivery and Tendon Embodiments 5

Tissue is generally not recognized to have the capability to delivergrowth factors (GF). GF delivery can be important to retain the GF atthe site of repair for sufficient time to allow repair/regeneration.Additionally, the repair process is a temporal process and the additionof GF over time can aid different stages of the repair process. Incontrast, a bolus delivery of GF may have a short-term effect or mayhave no or minimal effect because the GF are rapidly removed from therepair site and cannot influence downstream stages of the repairprocess.

Platelet rich plasma (PRP) is derived from centrifuging whole blood, hasa platelet concentration higher than that of whole blood, is thecellular component of plasma that settles after centrifugation, andcontains numerous growth factors (such as vascular endothelial growthfactor, transforming growth factor-β1 , and platelet-derived growthfactor-BB). PRP can be used in sports medicine by directly applying thePRP to the injury site. However, there is a lack of evidence showingthat PRP can facilitate the repair process. One potential explanationfor this is that PRP may not be adequately retained or delivered overthe appropriate time frame to permit biological activity. To circumventthis, some practitioners resort to multiple injections, although theobvious downside is multiple patient visits, incurred health care costsand pain/morbidity of the injection site.

Embodiments of the present invention combines the ground tendon aspreviously described with PRP. Appropriate processing of the groundtendon is beneficial to retain the physical properties of the collagenand extracellular matrix so as to allow slow activation of platelets andthe retention of the growth factors that are released from the plateletson the tendon extracellular matrix. These growth factors may interact byhydrophobic, van der walls, ionic, and other means to bind to the ECM.

Controlled delivery of PRP has been previously described using a gelatincarrier (denatured collagen hydrogel). However, ground tendon moreclosely resembles the intended repair structure and may be preferred.Further, without being bound by any particular theory, it is believedthat intact collagen and extracellular matrix can retain the ECMstructures better to retain the GF. Exemplary embodiments of the instantinvention encompass tendon and PRP compositions that provide for timedand/or controlled release of PRP therefrom, for a variety of tissuerepair processes, including cartilage, bone, tendon, and the like.

According to embodiments of the present invention, it is possible to useautologous PRP rather than recombinant growth factors. Additionally, thePRP may be derived from an allogeneic source. In this case the PRP andground tendon can be combined during the manufacturing process andlyophilized. Lyophilization can control the porosity such as to create aphysical barrier to growth factor release. Such porosity could becontrolled by methods such as freezing rate, NaCl concentration, and thelike. Other allogeneic growth factors can be added to the ground tendonsuch as BMP-2 , -4 and -7 and TGFb-1 extracted from demineralized boneto allow for temporal release of a different set of GF.

There are a number of medical applications. These include the repair ofpartial tendon tears in which the ground tendon/PRP composite can bedelivered to the tear area to facilitate repair. The product compositecan also be delivered to facilitate repair of meniscal tears. Typically,the product composite can be delivered to the repair site and the repaircan be closed using bioresorbable pins, tacks or sutures. The productcomposite may also be applied to tendon—bone interface such as forrotator cuff repair. The product composite can be delivered to theinterface or footprint area prior to the sutures being pulled tight toposition the tendon and bone together with the ground tendon/PRP at theinterface. Postoperative rotator cuff tears occur from 11-94% of rotatorcuff surgeries. Because of the high retear rate, it is beneficial toexplore techniques of biological augmentation of rotator cuff repair.The normal tendon to bone interface includes tendon, non-mineralizedfibrocartilage, mineralized fibrocartilage and bone. The normal repairprocess produces substantial fibrovascular/scar tissue which leads toinferior repair zone and substandard biomechanical strength. Appropriatedelivery of the PRP with the appropriate scaffold (ground tendon) canfacilitate the appropriate tissue structure.

Hence, embodiments of the present invention encompass compositions andmethods that involve flowable/ground tendon (e.g. allograft tissue),optionally for use in delivering drugs, growth factors, and the like.

Bone (or Bone with Mesenchymal Stem Cell) Embodiments

Tissue matrix compositions as disclosed herein, such as soft tissuematrix compositions and the like, may also include bone tissue material,optionally in combination with mesenchymal stem cells. For example,treatment compositions and methods may encompass the use of a flowabledecellularized or de-epidermalized skin component (or other soft tissuematrix), in combination with a demineralized bone matrix (DBM)component. A DBM component may include, for example, bone (e.g.allograft) from which inorganic mineral has been removed. In someinstances, a DBM component may include an organic collagen matrixmaterial. Relatedly, treatment compositions and methods may encompassthe use of a flowable decellularized or de-epidermalized skin component(or other soft tissue matrix), in combination with a material thatincludes both a bone material component and a mesenchymal stem cellcomponent. For example, a flowable skin or soft tissue matrix materialmay include an amount of ALLOSTEM® Stem Cell Bone Growth Substitute.Relatedly, a flowable skin or soft tissue matrix material may include anamount of adipose-derived mesenchymal stem cells combined with partiallydemineralized cancellous bone. In some cases, the bone material and/ormesenchymal stem cells may be present in a morselized form. Hence,compositions and methods as disclosed herein may include a flowable softtissue or skin matrix material combined with ALLOSTEM® morsels, so as toform a bone putty. Exemplary bone and mesenchymal stem cell compositionsand methods for their preparation are described in US 2010/0124776 toShi, the contents of which are incorporated herein by reference.

According to some embodiments, flowable decellularized skin orde-epidermilized skin (or other soft tissue) can be mixed or combinedwith DBM morsels or ALLOSTEM® morsels (adipose-derived mesenchymal stemcells combined with partially demineralized cancellous bone) to make aputty formulation with good handling characteristics. For example, suchmorsels or putty compositions may stay in place upon implantation.Relatedly, such morsels or putty compositions may persist at the site ofapplication (e.g. bony defect area) and resist removal by irrigationand/or contact with blood. In some instances, flowable decellularizedskin or de-epidermilized skin (or other soft tissue) can provide aneffective carrier to hold DBM and/or mesenchymal stem cells in place andprevent their migration. Such carriers may provide enhanced performanceover other carriers such as hyaluronic acid, chitosan, pluronic acid,ceramic cements, carboxymethylcellulose, calcium sulfate, which aretypically synthetic, not derived from human tissue, or otherwise do notinclude flowable decellularized skin or de-epidermilized skin (or othersoft tissue). Further, such morsels or putty compositions may remainsufficiently moist or hydrated, such that they are not too dry orcrumbly. What is more, such morsels or putty compositions can beterminally sterilized at 15-35 kGy by e-beam for example, withoutcompromising the handling characteristics, such that the putty can bemolded into any desired shape, without being or becoming too thin orrunny.

While exemplary embodiments have been described in some detail, by wayof example and for clarity of understanding, those of skill in the artwill recognize that a variety of modification, adaptations, and changesmay be employed. Hence, the scope of the present invention should belimited solely by the claims.

What is claimed is:
 1. A soft tissue matrix composition for use in apatient treatment, the composition consisting of a cryofractured humansoft tissue material having a mechanically disrupted extracellularmatrix collagen or macrostructure and a partially disrupted collagenmicrofibrillar architecture, wherein the cryofractured tissue materialis in a naturally hydrated state, the hydration comprising the naturalinterstitial water of the cryofractured soft tissue material that ispresent prior to cryofractionation.
 2. The soft tissue matrixcomposition according to claim 1, wherein the soft tissue matrixcomposition is present in an applicator assembly.
 3. The soft tissuematrix composition according to claim 1, wherein the human soft tissuematerial is acellular.
 4. The soft tissue matrix composition accordingto claim 1, wherein the human soft tissue material is at least partiallydecellularized.
 5. The soft tissue matrix composition according to claim1, wherein the human soft tissue material is not partially or completelydecellularized.
 6. The soft tissue matrix composition according to claim1, wherein the soft tissue matrix composition does not adhere to asurgical glove material.
 7. The soft tissue matrix composition accordingto claim 1, wherein the soft tissue matrix composition does not adhereto a material selected from the group consisting of latex, neoprene,vinyl, and nitrile.
 8. The soft tissue matrix composition according toclaim 1, wherein the soft tissue matrix composition has a puttyconsistency.
 9. The soft tissue matrix composition according to claim 1,wherein the soft tissue matrix composition has a shear resistance of250-2,500 Pascal seconds at room temperature.
 10. The soft tissue matrixcomposition according to claim 1, wherein the soft tissue matrixcomposition has a shear resistance of 1,000-2,000 Pascal seconds at roomtemperature.
 11. The soft tissue matrix composition according to claim1, wherein the soft tissue matrix composition can pass through a borehole of a 18 g needle and not through a bore hole of a 26 g needle. 12.The soft tissue matrix composition according to claim 1, wherein atleast a portion of the human soft tissue material is in minced or slicedpieces.
 13. The soft tissue matrix composition according to claim 1,wherein at least a portion of the soft tissue matrix composition is incomminuted form.
 14. The soft tissue matrix composition according toclaim 1, wherein the human soft tissue material is selected from thegroup consisting of amnion tissue, umbilical cord tissue, chorionictissue, intestine tissue, tendon tissue, and skin tissue.
 15. The softtissue matrix composition according to claim 1, wherein the human softtissue material is skin.
 16. The soft tissue matrix compositionaccording to claim 1, wherein the human soft tissue material is fullthickness skin or skin lacking the epidermal layer.
 17. The soft tissuematrix composition according to claim 1, wherein the human soft tissuematerial is amnion tissue.
 18. The soft tissue matrix compositionaccording to claim 1, wherein the human soft tissue material isumbilical cord tissue.
 19. The soft tissue matrix composition accordingto claim 1, wherein the human soft tissue material is chorionic tissue.20. The soft tissue matrix composition according to claim 1, wherein thehuman soft tissue material is intestine tissue.
 21. The soft tissuematrix composition according to claim 1, wherein the human soft tissuematerial is tendon tissue.
 22. A soft tissue matrix composition for usein a patient treatment, the composition consisting of a cryofracturedand triturated human soft tissue material having a mechanicallydisrupted extracellular matrix collagen or macrostructure and apartially disrupted collagen microfibrillar architecture, wherein thecryofractured and triturated tissue material is in a naturally hydratedstate, the hydration comprising the natural interstitial water of thecryofractured and triturated human soft tissue material that is presentprior to cryofractionation and trituration.
 23. The soft tissue matrixcomposition according to claim 22, wherein the soft tissue matrixcomposition is present in an applicator assembly.
 24. The soft tissuematrix composition according to claim 22, wherein the human soft tissuematerial is acellular.
 25. The soft tissue matrix composition accordingto claim 22, wherein the human soft tissue material is at leastpartially decellularized.
 26. The soft tissue matrix compositionaccording to claim 22, wherein the human soft tissue material is notpartially or completely decellularized.
 27. The soft tissue matrixcomposition according to claim 22, wherein the soft tissue matrixcomposition does not adhere to a surgical glove material.
 28. The softtissue matrix composition according to claim 22, wherein the soft tissuematrix composition does not adhere to a material selected from the groupconsisting of latex, neoprene, vinyl, and nitrile.
 29. The soft tissuematrix composition according to claim 22, wherein the soft tissue matrixcomposition has a putty consistency.
 30. The soft tissue matrixcomposition according to claim 22, wherein the soft tissue matrixcomposition has a shear resistance of 250-2,500 Pascal seconds at roomtemperature.
 31. The soft tissue matrix composition according to claim22, wherein the soft tissue matrix composition has a shear resistance of1,000-2,000 Pascal seconds at room temperature.
 32. The soft tissuematrix composition according to claim 22, wherein the soft tissue matrixcomposition can pass through a bore hole of a 18 g needle and notthrough a bore hole of a 26 g needle.
 33. The soft tissue matrixcomposition according to claim 22, wherein at least a portion of thehuman soft tissue material is in minced or sliced pieces.
 34. The softtissue matrix composition according to claim 22, wherein at least aportion of the soft tissue matrix composition is in comminuted form. 35.The soft tissue matrix composition according to claim 22, wherein thehuman soft tissue material is selected from the group consisting ofamnion tissue, umbilical cord tissue, chorionic tissue, intestinetissue, tendon tissue, and skin tissue.
 36. The soft tissue matrixcomposition according to claim 22, wherein the human soft tissuematerial is skin.
 37. The soft tissue matrix composition according toclaim 22, wherein the human soft tissue material is full thickness skinor skin lacking the epidermal layer.
 38. The soft tissue matrixcomposition according to claim 22, wherein the human soft tissuematerial is amnion tissue.
 39. The soft tissue matrix compositionaccording to claim 22, wherein the human soft tissue material isumbilical cord tissue.
 40. The soft tissue matrix composition accordingto claim 22, wherein the human soft tissue material is chorionic tissue.41. The soft tissue matrix composition according to claim 22, whereinthe human soft tissue material is intestine tissue.
 42. The soft tissuematrix composition according to claim 22, wherein the human soft tissuematerial is tendon tissue.
 43. A soft tissue matrix composition for usein a patient treatment prepared by a method consisting of: providing ahuman soft tissue material; processing the human soft tissue material tomechanically disrupt the extracellular matrix collagen or macrostructureand a partially disrupt the collagen microfibrillar architecture by aprocess selected from the group consisting of: (i) cryofracturing thehuman soft tissue material; (ii) cryofracturing and washing the humansoft tissue material; (iii) cryofracturing and triturating the humansoft tissue material; and (iv) cryofracturing, triturating, and washingthe human soft tissue material, wherein the processed tissue material isin a naturally hydrated state, the hydration comprising a substantialamount of the natural interstitial water of the human soft tissuematerial.
 44. The soft tissue matrix composition according to claim 43,wherein the soft tissue matrix composition is present in an applicatorassembly.
 45. The soft tissue matrix composition according to claim 43,wherein the human soft tissue material is acellular.
 46. The soft tissuematrix composition according to claim 43, wherein the human soft tissuematerial is at least partially decellularized.
 47. The soft tissuematrix composition according to claim 43, wherein the human soft tissuematerial is not partially or completely decellularized.
 48. The softtissue matrix composition according to claim 43, wherein the soft tissuematrix composition does not adhere to a surgical glove material.
 49. Thesoft tissue matrix composition according to claim 43, wherein the softtissue matrix composition does not adhere to a material selected fromthe group consisting of latex, neoprene, vinyl, and nitrile.
 50. Thesoft tissue matrix composition according to claim 43, wherein the softtissue matrix composition has a putty consistency.
 51. The soft tissuematrix composition according to claim 43, wherein the soft tissue matrixcomposition has a shear resistance of 250-2,500 Pascal seconds at roomtemperature.
 52. The soft tissue matrix composition according to claim43, wherein the soft tissue matrix composition has a shear resistance of1,000-2,000 Pascal seconds at room temperature.
 53. The soft tissuematrix composition according to claim 43, wherein the soft tissue matrixcomposition can pass through a bore hole of a 18 g needle and notthrough a bore hole of a 26 g needle.
 54. The soft tissue matrixcomposition according to claim 43, wherein at least a portion of thehuman soft tissue material is in minced or sliced pieces.
 55. The softtissue matrix composition according to claim 43, wherein at least aportion of the soft tissue matrix composition is in comminuted form. 56.The soft tissue matrix composition according to claim 43, wherein thehuman soft tissue material is selected from the group consisting ofamnion tissue, umbilical cord tissue, chorionic tissue, intestinetissue, tendon tissue, and skin tissue.
 57. The soft tissue matrixcomposition according to claim 43, wherein the human soft tissuematerial is skin.
 58. The soft tissue matrix composition according toclaim 43, wherein the human soft tissue material is full thickness skinor skin lacking the epidermal layer.
 59. The soft tissue matrixcomposition according to claim 43, wherein the human soft tissuematerial is amnion tissue.
 60. The soft tissue matrix compositionaccording to claim 43, wherein the human soft tissue material isumbilical cord tissue.
 61. The soft tissue matrix composition accordingto claim 43, wherein the human soft tissue material is chorionic tissue.62. The soft tissue matrix composition according to claim 43, whereinthe human soft tissue material is intestine tissue.
 63. The soft tissuematrix composition according to claim 43, wherein the human soft tissuematerial is tendon tissue.
 64. The soft tissue matrix compositionaccording to claim 43, wherein the processing is cryofracturing thehuman soft tissue material.
 65. The soft tissue matrix compositionaccording to claim 43, wherein the processing is cryofracturing andwashing the human soft tissue material.
 66. The soft tissue matrixcomposition according to claim 43, wherein the processing iscryofracturing and triturating the human soft tissue material.
 67. Thesoft tissue matrix composition according to claim 43, wherein theprocessing is cryofracturing, triturating, and washing the human softtissue material.